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Identification and Functional Validation of a Biomarker for the Diagnosis of Miltefosine Relapse during Visceral Leishmaniasis

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  • 1 Department of Medicine, Infectious Disease Research Laboratory, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India
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Miltefosine is the only orally administrable drug for the treatment of leishmaniasis. But in recent years, a decline in its efficacy points toward the emergence of resistance to this drug. Knowledge of biomarkers for miltefosine resistance may be beneficial for proper selection of treatment regimen. Splenic aspirates were collected and parasites cultured from patients relapsed after initial cure (N = 15) and successfully treated (N = 15) with miltefosine. Differential expression of genes in miltefosine-resistant strains was examined by DNA microarray and validated by real-time reverse transcription polymerase chain reaction and Western blotting. Of 669 upregulated genes, the cysteine protease-like protein of calpain family (GenBank: CBZ34784) was found to be significantly overexpressed in resistant parasite strains and only anti-calpain antibodies showed its presence in the sera of relapse patients through Western blotting. Calpain family cysteine protease-like protein can be useful as a potential biomarker of miltefosine unresponsiveness.

Author Notes

Address correspondence to Shyam Sundar, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India. E-mail: drshyamsundar@hotmail.com

Financial support: This research was funded by a grant from the National Institute of Allergy and Infectious Diseases, NIH, USA, vide grant no. P50AI074321. Puja Tiwary and Dinesh Kumar thank the Council of Scientific and Industrial Research (CSIR) for providing fellowships.

Authors’ addresses: Puja Tiwary, Dinesh Kumar, and Shyam Sundar, Department of Medicine, Infectious Disease Research Laboratory, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India, E-mails: tiwarypuja@gmail.com, praja_dinesh@yahoo.co.in, and drshyamsundar@hotmail.com.

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